NAR患者鼻灌洗EOS与诱导痰EOS的相关性研究

目的:评估非变应性鼻炎(nonallergic rhinitis,NAR)患者全身、鼻腔及下气道嗜酸性粒细胞(Eosinophils,EOS)浸润水平及相关性。探讨EOS在NAR患者鼻腔及下气道的特征及意义。方法:2011年6月~2012年6月在我院就诊的NAR患者241例,同期征集健康对照组222例,所有受试者均进行病史采集、皮肤点刺实验(SPT)、血清嗜酸性粒细胞、鼻灌洗、诱导痰检查。结果:1NAR组和对照组相比鼻灌洗EOS计数、诱导痰EOS比例、血EOS比例均显著高于健康对照组(P均0.01)。2有鼻腔EOS炎症和无鼻腔EOS炎症层的诱导痰EOS百分比阳性率分别为34.7%vs 9.6%(P0.01)。3NAR组鼻灌洗EOS计数和诱导痰EOS比例存在明显相关性(r=0.262,P=0.000),鼻灌洗EOS计数和血EOS比例存在明显相关性(r=0.228,P=0.000),诱导痰EOS比例和血EOS比例存在明显相关性(r=0.291,P=0.000)。结论:NAR患者血液、鼻腔及下气道EOS浸润程度较正常对照组均明显升高,EOS在NAR患者血液、鼻腔及下气道炎症中存在明显一致性,提示NAR是一种全身系统性炎症性疾病;无下气道症状的NAR患者部分存在下气道EOS炎症,鼻腔及血液EOS炎症是导致下气道EOS炎症的主要高危因素。鼻灌洗EOS可能是观测NAR患者下气道炎症及对下气道炎症进行评估和追踪的有效指标。EOS可能是鼻炎、哮喘及血液炎症相关的效应细胞,是上下气道炎症有效生物标记物。     

呼出气一氧化氮检测在哮喘-慢阻肺重叠综合征中的临床应用

目的:探讨哮喘-慢性阻塞性肺疾病重叠综合征(asthma-chronic obstructive pulmonary disease overlap syndrome,ACOS)患者呼出气一氧化氮浓度(FENO)的变化情况及与一秒用力呼气容积(FEV1)、血嗜酸性粒细胞(eosnophil,EOS)及超敏C反应蛋白(high sensitivity C reactive protein,hs-CRP)的关系。方法:选取上海第九人民医院2016年1月~2016年12月呼吸内科病区收治的ACOS患者30例及无肺部疾病的健康的体检人群20例分别作为实验组和对照组。治疗前后分别检测和比较两组FENO、FEV1值、EOS及血清hs-CRP水平的变化。结果:治疗前,实验组FENO、hs-CRP及EOS水平显著高于对照组(P值分别为0.0081,0.0263及0.0078),FEV1%水平显著低于对照组(P=0.0047)。治疗后,两组间FENO、hs-CRP及EOS水平变化无显著差异(P值分别为0.2614,0.1347及0.2431),而实验组FEV1%水平仍显著低于对照组(P=0.0469)。实验组在治疗前后FENO、hs-CRP及EOS水平均显著下降(P值分别为0.0027,0.0427,0.0031),而FEV1%变化无显著差异(P=0.1427)。在治疗前后,FENO水平与hs-CRP水平呈正相关(γ=0.7392,P=0.0168;γ=0.7214,P=0.0248),与血EOS水平亦呈正相关(γ=0.8782,P=0.0072;γ=0.7642,P=0.0231)。而FENO水平与FEV1%在治疗前后无相关性(P0.05)。结论:ACOS患者气道内存在嗜酸性粒细胞相关性慢性炎症,短期静脉糖皮质激素治疗可缓解其气道内炎症。ACOS患者FENO水平与血EOS、血清hs-CRP水平有显著相关性,可用于辅助评估ACOS患者静脉糖皮质激素疗效。     

痰嗜酸性粒细胞检测与AECOPD全身激素使用关系

为探讨痰嗜酸性粒细胞(EOS)对慢性阻塞性肺病(COPD)急性加重全身激素治疗的指导作用。本研究选取AECOPD住院的58例患者,根据诱导痰细胞分类将痰EOS≥3%定为A组(18例),EOS3%随机分为B组(20例)、C组(20例)。三组都进行抗生素、支气管扩张剂治疗,此外A组、B组再接受强的松片口服治疗2周,三组患者在治疗前、治疗后2周分别进行肺功能测定、痰细胞计数、慢阻肺患者自我评估测试(CAT问卷)和CRP测定。并随访4周,观察治疗失败病例。通过相关信息表明,治疗前A组CAT评分、支气管舒张试验阳性率高于B组、C组,痰中性粒细胞、CRP低于B组、C组;治疗前后三组中在肺功能、CAT量表改善A组效果最为显著,B组效果最差。研究表明痰嗜酸性粒细胞检查可以对COPD急性加重患者进行分层,作为优化激素治疗的指标。     

过敏性紫癜兔模型的免疫学改变及机制初探

目的通过对过敏性紫癜兔模型免疫学改变的初步研究,探讨该病的免疫学发病机制。方法通过对过敏性紫癜兔模型进行血常规检测、ELISA方法检测免疫细胞及细胞因子CD3、CD4、CD8、CD4/CD8、IL-2、TNF-α含量;免疫荧光方法检测皮肤、肾脏免疫球蛋白IgA,IgG及补体C3;肾小球Masson染色、PAS染色;皮肤、肺组织的Luna染色等,并与对照组进行比较分析。结果与对照组相比,模型组兔血白细胞（WBC）增多,中性粒细胞（NEU）及百分比增高,嗜酸性粒细胞（EOS）及百分比增高,嗜碱性粒细胞增多等,差别均具有统计学意义;外周血CD3＋T淋巴细胞含量减少,CD4＋T淋巴细胞含量减少,CD8＋T淋巴细胞含量增多,CD4＋/CD8＋比值下降,细胞因子IL-2水平下降,TNF-α水平升高,差别均具有统计学意义;皮肤、肾脏免疫球蛋白IgA、IgG、C3表达增多;肾小球胶原纤维增生,系膜增厚,系膜基质增多;皮肤真皮层及肺组织内嗜酸性粒细胞表达增多。结论将为明确其发病机制,临床诊断和疗效观察找出新的指标,选择适当的治疗方案提供有价值的理论依据。     

嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断及预后预测价值分析

摘要 目的：探讨嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断及预后预测价值。方法：选取我院2020年3月到2023年3月收治的100例慢性鼻窦炎伴鼻息肉患者作为研究对象,将其分为慢性鼻窦炎伴鼻息肉组,选取同期来我院治疗的100例单纯慢性鼻窦炎患者作为慢性鼻窦炎组,另选取同期来我院体检的100名健康志愿者作为对照组。对比三组受检者嗜酸性粒细胞、调节性T细胞表达水平,并建立受试者工作特征（ROC）曲线,分析嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断效能。随后对100例性鼻窦炎伴鼻息肉患者手术治疗后进行1年随访,依照患者的复发情况评价其预后情况,并分为两个亚组,将术后1年内复发的21例患者分为预后不良组,两未复发的79例患者分为预后良好组,对比两组患者一般临床情况,嗜酸性粒细胞、调节性T细胞表达水平,并分析嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的预后预测价值。结果：三组受检者嗜酸性粒细胞、调节性T细胞表达水平对比差异显著,慢性鼻窦炎伴鼻息肉组患者嗜酸性粒细胞（EOS）个数高于慢性鼻窦炎组和对照组,调节性T细胞（Tregs）水平低于慢性鼻窦炎组和对照组（P＜0.05）;通过绘制ROC曲线,确定EOS、Tregs其对慢性鼻窦炎伴鼻息肉的诊断效能,结果显示,EOS、Tregs两者联合对慢性鼻窦炎伴鼻息肉的诊断效能优于单一检测（P＜0.05）;预后良好组与预后不良组患者性别、年龄、BMI、吸烟史、饮酒史、白细胞介素-35（IL-35）、转化生长因子-β（TGF-β）、白细胞介素-1β（IL-1β）表达水平对比无明显差异（P＞0.05）,预后良好组与预后不良组患者病程、合并哮喘、合并变应性鼻炎、组织淋巴细胞占比、白细胞介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）、EOS个数以及Tregs表达水平对比差异显著（P＜0.05）;logistic回归分析结果表明：合并哮喘、组织淋巴细胞占比、EOS个数以及Tregs为慢性鼻窦炎伴鼻息肉的预后独立影响因素（P＜0.05）。结论：嗜酸性粒细胞、调节性T细胞不仅对慢性鼻窦炎伴鼻息肉的临床诊断具有重要价值,而且能够预测慢性鼻窦炎伴鼻息肉患者的预后情况,因此临床上对于EOS个数增加,Tregs降低的患者要及时改善治疗措施,预防患者预后不良的发生。     

糖皮质激素下调JAK/STAT抑制嗜酸粒细胞哮喘

摘要 目的：探究糖皮质激素对嗜酸粒细胞哮喘(Eosinophilic asthma, EA) 2 型固有免疫细胞(Type 2 innate lymphoid cells, ILC2s)的影响及相关机制。方法：研究对象来自我院 2021年6月至 2022年6月的EA患者和健康对照（Healthy control, HC）,收集相应临床基线资料并评估病情、进行血常规、肺功能等检查;应用流式细胞术检测外周血单个核细胞（Peripheral blood mononuclear cell, PBMC） ILC2s(CD45+Lin-CD127+CD294+);ELISA检测外周血IL-5、IL-13浓度。糖皮质激素治疗EA 患者3月后,观察PBMC中ILC2s及IL-5、IL-13浓度。C57BL/6J小鼠给予鸡卵清蛋白(Ovalbumin,OVA) 20 μg 腹腔注射致敏后用1%OVA雾化吸入激发哮喘EA模型,阴性对照（Negative control, NC）组小鼠用同等体积PBS作为对照。EA造模成功的小鼠通过流式细胞术检测血液及肺泡灌洗液中ILC2s,HE染色检测小鼠肺泡灌洗液中嗜酸性粒细胞(Eosinophil, EOS)及肺部炎症。EA小鼠经糖皮质激素处理后,检测肺部炎症情况;流式细胞术检测PBMC、肺泡灌洗液（Bronchoalveolar lavage fluid, BALF）中 ILC2s;分离肺组织ILC2s,western blot检测相关蛋白表达情况。结果：EA组的ILC2s比例升高, EOS升高,2型细胞因子IL-5、IL-13增加,糖皮质激素治疗1月及3月后ILC2s比例下降,2型细胞因子IL-5、IL-13下降。与NC组小鼠比较,EA组小鼠PBMC及BALF中ILC2s升高,BALF中EOS升高,血清中2型细胞因子IL-5、IL-13升高,肺部炎症加重。糖皮质激素治疗后,肺部炎症减轻,EOS下降,ILC2s减少,2型细胞因子IL-5、IL-13下降,下调JAK/STAT蛋白。结论：在EA中,糖皮质激素通过下调JAK/STAT蛋白抑制ILC2s的功能减轻肺部炎症,为激素治疗嗜酸性粒细胞哮喘的机制提供了新方向。     

老年慢性嗜酸性粒细胞肺炎一例

目的:总结1例高龄患有慢性嗜酸性粒细胞肺炎(CEP)的诊断和治疗的临床过程,探讨最佳的治愈方法。方法:对1例患有慢性嗜酸性粒细胞肺炎的患者进行详细检查、诊治,并结合文献资料进行分析,对其病症的临床症状和诊断治疗予以讨论。结果:CEP的病因不是很明确,患者多以以往有过敏病史的临床特点,同时也比较容易复发;但是经过治疗,有效地减少了复发,治愈效果良好。结论:CEP具有其特有的典型症状,经过给予激素维持治疗可明显改善病症,控制病情的发展变化。     

茎点霉致泛发性嗜酸性脓疱性毛囊炎1例及实验分析

目的报道1例顽固性嗜酸性脓疱性毛囊炎,并对其病因、诊断及治疗方法进行研究讨论。方法患者男性,47岁,头面部丘疱疹1年伴瘙痒,泛发,并出现血嗜酸性粒细胞增多3个月就诊。取皮损组织和骨髓进行组织分别进行HE、PAS、PASM、钙荧光白染色;同时进行组织真菌培养;真菌特异性引物ITS4/ITS5、NL1/NL5、T1/TUB4Rd测序。结果组织病理可见以毛囊为中心的嗜酸性毛囊化脓性炎,PAS染色和钙荧光白染色可见真皮、皮下组织大量不规则菌丝和孢子,毛囊及毛囊周围显著;骨髓PASM染色可见大量芽生孢子,组织及骨髓组织培养见绒毛状菌落,初期为粉红色,渐变为黑褐色,培养4周见黑色子囊形成;序列鉴定为茎点霉属;抗真菌治疗后痊愈。结论嗜酸性脓疱性毛囊炎可由真菌感染诱发;作为植物致病菌的茎点霉属感染可导致严重的变态反应,并可诱发嗜酸性脓疱性毛囊炎。     

孟鲁司特钠联合左卡巴斯汀鼻喷剂治疗小儿过敏性鼻炎的疗效评价及对血清ECP、EOS及CRP水平的影响

目的:探讨孟鲁司特钠联合左卡巴斯汀鼻喷剂治疗小儿过敏性鼻炎的疗效及对血清嗜酸性粒细胞阳离子蛋白(ECP)、嗜酸性粒细胞(EOS)及C反应蛋白(CRP)水平的影响。方法:选择我院2016年7月～2018年7月收治的151例过敏性鼻炎患儿,按随机数字表法分为69例对照组和82例观察组,对照组采用左卡巴斯汀鼻喷剂治疗,观察组在对照组基础上联合孟鲁司特钠治疗,比较两组临床疗效,治疗前后症状及体征评分,生活质量评分,血清ECP、EOS及CRP、总免疫球蛋白E(TIg E)和特异性免疫球蛋白E(Sig E)水平,和不良反应发生情况。结果:观察组总有效率高于对照组,差异有统计学意义(P0.05)。治疗前,两组症状及体征评分,生活质量评分,血清ECP、EOS及CRP、总免疫球蛋白E(TIg E)和特异性免疫球蛋白E(Sig E)水平比较差异无统计学意义(P0.05);治疗后,两组以上指标均下降,观察组低于对照组,差异均有统计学意义(均P0.05)。两组均有胃肠道反应、口干、头痛发生,组间总不良反应发生率比较无统计学差异(P0.05)。结论:孟鲁司特钠联合左卡巴斯汀鼻喷剂治疗小儿过敏性鼻炎安全有效,能够降低血清ECP、EOS及CRP水平,促进患儿恢复。     

孟鲁司特钠联合布地奈德治疗对小儿慢性咳嗽患者IgE、ECP、EOS水平的影响

目的探讨孟鲁司特钠联合布地奈德治疗对小儿慢性咳嗽治疗效果及患儿血清免疫球蛋白(IgE)、血清嗜酸性粒细胞阳离子蛋白(ECP)和外周血嗜酸性粒细胞计数(EOS)的影响。方法选取90例慢性咳嗽患儿随机分为治疗组和对照组,每组45例。两组患者均给予化痰止咳、抗生素等常规治疗。对照组给予布地奈德气雾剂进行吸入治疗,治疗组在对照组的基础上给予口服孟鲁司特钠,两组均持续治疗4周。观察两组患儿治疗前后IgE、ECP、EOS水平变化,并比较其临床疗效及不良反应。结果治疗4周后,两组患儿IgE、ECP、EOS均显著下降,治疗组下降水平大于对照组(267.25±35.12 vs 429.56±59.12,6.12±1.38 vs 10.52±3.81,0.39±0.05 vs 0.62±0.09;t=-15.8338,t=-7.2839,t=-14.9858;P0.01);治疗组患儿总有效率明显高于对照组(91.1%vs 71.1%,χ2=7.42,P0.05);治疗过程中两组均无严重的不良反应。结论孟鲁司特钠联合布地奈德治疗治疗小儿慢性咳嗽疗效比单独使用布地奈德有明显增强,可能与降低血清IgE、ECP、EOS水平有关。     

Analysing the eosinophil cationic protein - a clue to the function of the eosinophil granulocyte

Eosinophil granulocytes reside in respiratory mucosa including lungs, in the gastro-intestinal tract, and in lymphocyte associated organs, the thymus, lymph nodes and the spleen. In parasitic infections, atopic diseases such as atopic dermatitis and asthma, the numbers of the circulating eosinophils are frequently elevated. In conditions such as Hypereosinophilic Syndrome (HES) circulating eosinophil levels are even further raised. Although, eosinophils were identified more than hundred years ago, their roles in homeostasis and in disease still remain unclear. The most prominent feature of the eosinophils are their large secondary granules, each containing four basic proteins, the best known being the eosinophil cationic protein (ECP). This protein has been developed as a marker for eosinophilic disease and quantified in biological fluids including serum, bronchoalveolar lavage and nasal secretions. Elevated ECP levels are found in T helper lymphocyte type 2 (atopic) diseases such as allergic asthma and allergic rhinitis but also occasionally in other diseases such as bacterial sinusitis. ECP is a ribonuclease which has been attributed with cytotoxic, neurotoxic, fibrosis promoting and immune-regulatory functions. ECP regulates mucosal and immune cells and may directly act against helminth, bacterial and viral infections. The levels of ECP measured in disease in combination with the catalogue of known functions of the protein and its polymorphisms presented here will build a foundation for further speculations of the role of ECP, and ultimately the role of the eosinophil.     

Deposition of eosinophil cationic protein in granulomas in allergic granulomatosis and vasculitis: the Churg-Strauss syndrome.

Biopsy specimens and tissues obtained at necropsy from two women who died after developing the Churg-Strauss syndrome were analysed to see whether granulomas in these patients contained activated eosinophils or secreted eosinophil cationic proteins, or both. Immunocytochemical studies with monoclonal antibody EG2 showed large amounts of eosinophil cationic protein and eosinophil protein-X (which are toxic for heart cells and other tissues) in the granulomas. Many activated and degranulating eosinophils were seen to be migrating from the blood into these areas. Eosinophils may play a central part in the development of lesions in the heart and other tissues in the Churg-Strauss syndrome.     

慢性腹痛患儿C~(13)-尿素呼气试验与胃镜检查的临床意义

目的了解慢性腹痛患儿幽门螺杆菌(H.pylori)的感染状态及幽门螺杆菌感染患儿内镜下表现的特点。方法应用C13尿素呼气试验,对905例以慢性腹痛为主要症状的患儿进行检测,对C13呼气试验阳性者进行电子胃镜检查。结果905例慢性腹痛患儿中H.pylori呈阳性185例(20.44%),随年龄增长,其H.pylori阳性率升高,学年组已达高峰。对H.pylori阳性者进行胃镜检查结果显示十二指肠隆起病变47例占25.40%,结节性胃炎41例占22.1%,慢性浅表性`胃炎38例占20.5%,结节性胃炎伴十二指肠隆起病变23例占12.43%,十二指肠球部溃疡23例占12.4%。胃溃疡7例,占3.7%(其中包括1例复合性溃疡),结节性胃炎伴十二指肠炎6例,占3.2%。结论H.pylori感染为小儿慢性腹痛的主要原因之一,也是导致慢性胃炎及消化性溃疡的主要原因之一。C13尿素呼气试验方便,快速,无痛苦,无放射性,是一较好的H.pylori检测方法;对既有消化道症状同时C13呼气试验阳性者进行胃镜检查能够协助临床诊断及治疗。     

Regulation of eosinophilopoiesis in a murine model of asthma

Eosinophilic inflammation plays a key role in tissue damage that characterizes asthma. Eosinophils are produced in bone marrow and recent observations in both mice and humans suggest that allergen exposure results in increased output of eosinophils from hemopoietic tissue in individuals with asthma. However, specific mechanisms that alter eosinophilopoiesis in this disease are poorly understood. The current study used a well-characterized murine animal model of asthma to evaluate alterations of eosinophil and eosinophil progenitor cells (CFU-eo) in mice during initial sensitization to allergen and to determine whether observed changes in either cell population were regulated by T lymphocytes. Following the first intranasal installation of OVA, we observed sequential temporal elevation of eosinophils in bone marrow, blood, and lung. In immunocompetent BALB/c mice, elevation of bone marrow eosinophils was accompanied by transient depletion of CFU-eo in that tissue. CFU-eo rebounded to elevated numbers before returning to normal baseline values following intranasal OVA exposure. In T cell-deficient BALB/c nude (BALB/c(nu/nu)) mice, CFU-eo were markedly elevated following allergen sensitization, in the absence of bone marrow or peripheral blood eosinophilia. These data suggest that eosinophilia of asthma results from alterations in two distinct hemopoietic regulatory mechanisms. Elevation of eosinophil progenitor cells in the bone marrow is T cell independent and likely results from altered bone marrow stromal cell function. Differentiation of eosinophil progenitor cells and phenotypic eosinophilia is T cell dependent and does not occur in athymic nude mice exposed to intranasal allergen.     

Differential expression and activation of Rab27A in human eosinophils: relationship to blood eosinophilia

Eosinophil degranulation is thought to play a pathophysiological role in asthma. Rab27A is a GTP-binding protein that is known to be essential for the degranulation of several leukocyte subsets and thus may be essential for eosinophil granule exocytosis. Here, we show that Rab27A mRNA and protein are expressed in human eosinophils. We have developed a novel assay to assess Rab27A activation and have found a similar activation pattern of this protein upon stimulation of eosinophils, neutrophils and NK cells suggesting a similar function in these cell types. Interestingly, Rab27A expression was elevated in eosinophils from asthmatic donors. Furthermore, eosinophils from eosinophilic donors displayed more rapid Rab27A activation kinetics than those from donors with lower eosinophil counts. Given that elevated blood eosinophil numbers correlate with increased priming of eosinophils, this pattern of Rab27A activation suggests differential protein expression in activated cells may allow eosinophils to degranulate more rapidly upon stimulation.     

Role of inflammatory cells in Chagas' disease. III. Kinetics of human eosinophil activation upon interaction with parasites (Trypanosoma cruzi)

The kinetics of human eosinophil activation and granule secretion initiated by interaction with Trypanosoma cruzi amastigotes was studied by using a monoclonal IgG1 antibody (termed EG2) that is specific for an epitope present only in the secreted forms of both eosinophil cationic protein (ECP) and the eosinophil protein X (EP-X), and hence not detectable in unstimulated resting eosinophils. Studies were carried out by using electron microscopy and indirect immunofluorescence. In the electron microscopy studies, deposits of protein A-gold particles in parasite-containing eosinophils that had been incubated previously with EG2 antibody were first detected 4 hr after initiation of the eosinophil-amastigote interaction. Control tests performed with a monoclonal IgG1 unreactive with eosinophils showed no deposition of protein A-gold particles. EG2 antibody binding was confined to the crystalloid granule matrix, where ECP and EP-X are known to be stored. A similar kinetic pattern of ECP/EP-X solubilization and secretion was confirmed by the results of the indirect immunofluorescence experiments also showing the binding of EG2 antibody after 4 hr of cell-parasite interaction. The kinetics of ECP/EP-X solubilization and secretion paralleled the kinetics of destruction of internalized amastigotes, suggesting a role for these basic proteins in parasite killing. Consistent with this notion was the detection of ECP/EP-X in the fluid of phagocytic vacuoles containing amastigotes and associated with the ingested organisms at the same time as the parasites began to show structural alterations. These results outlined the kinetics of eosinophil activation in terms of the time required for mobilization of two basic proteins associated with eosinophil secretion that are known to be biologically active.     

Intestinal macrophage/epithelial cell-derived CCL11/eotaxin-1 mediates eosinophil recruitment and function in pediatric ulcerative colitis

Clinical studies have demonstrated a link between the eosinophil-selective chemokines, eotaxins (eotaxin-1/CCL11 and eotaxin-2/CCL24), eosinophils, and the inflammatory bowel diseases, Crohn's disease and ulcerative colitis (UC). However, the cellular source and individual contribution of the eotaxins to colonic eosinophilic accumulation in inflammatory bowel diseases remain unclear. In this study we demonstrate, by gene array and quantitative PCR, elevated levels of eotaxin-1 mRNA in the rectosigmoid colon of pediatric UC patients. We show that elevated levels of eotaxin-1 mRNA positively correlated with rectosigmoid eosinophil numbers. Further, colonic eosinophils appeared to be degranulating, and the levels positively correlated with disease severity. Using the dextran sodium sulfate (DSS)-induced intestinal epithelial injury model, we show that DSS treatment of mice strongly induced colonic eotaxin-1 and eotaxin-2 expression and eosinophil levels. Analysis of eosinophil-deficient mice defined an effector role for eosinophils in disease pathology. DSS treatment of eotaxin-2(-/-) and eotaxin-1/2(-/-) mice demonstrated that eosinophil recruitment was dependent on eotaxin-1. In situ and immunofluorescence analysis-identified eotaxin-1 expression was restricted to intestinal F4/80(+)CD11b(+) macrophages in DSS-induced epithelial injury and to CD68(+) intestinal macrophages and the basolateral compartment of intestinal epithelial cells in pediatric UC. These data demonstrate that intestinal macrophage and epithelial cell-derived eotaxin-1 plays a critical role in the regulation of eosinophil recruitment in colonic eosinophilic disease such as pediatric UC and provides a basis for targeting the eosinophil/eotaxin-1 axis in UC.     

Generation of Eosinophils from Cryopreserved Murine Bone Marrow Cells

Eosinophils are produced in the bone marrow from CD34⁺ eosinophil lineage–committed progenitors, whose levels in the bone marrow are elevated in a variety of human diseases. These findings suggest that increased eosinophil lineage–committed progenitor production is an important process in disease-associated eosinophilia. The pathways central to the biology of the eosinophil lineage–committed progenitor remain largely unknown. Thus, developing new methods to investigate the regulators of eosinophil lineage–committed progenitor differentiation is needed to identify potential therapeutic targets to specifically inhibit eosinophil production. We tested cytokine regimens to optimize liquid cultures for the study of eosinophil lineage–committed progenitor and eosinophil precursor differentiation into mature eosinophils. Stem cell factor (but not fms-related tyrosine kinase 3 ligand) was required for optimal yield of eosinophils. Furthermore, we evaluated the effects of cell preservation and scale on the culture, successfully culturing functional eosinophils from fresh and frozen murine bone marrow cells and in a standard-sized and 96-well culture format. In summary, we have developed an adaptable culture system that yields functionally competent eosinophils from murine low-density bone marrow cells and whose cytokine regime includes expansion of progenitors with stem cell factor alone with subsequent differentiation with interleukin 5.     

Decreased expression of membrane IL-5 receptor alpha on human eosinophils: I. Loss of membrane IL-5 receptor alpha on airway eosinophils and increased soluble IL-5 receptor alpha in the airway after allergen challenge

IL-5 is a key cytokine for eosinophil maturation, recruitment, activation, and possibly the development of inflammation in asthma. High concentrations of IL-5 are present in the airway after Ag challenge, but the responsiveness of airway eosinophils to IL-5 is not well characterized. The objectives of this study were to establish, following airway Ag challenge: 1) the expression of membrane (m)IL-5Ralpha on bronchoalveolar lavage (BAL) eosinophils; 2) the responsiveness of these cells to exogenous IL-5; and 3) the presence of soluble (s)IL-5Ralpha in BAL fluid. To accomplish these goals, blood and BAL eosinophils were obtained from atopic subjects 48 h after segmental bronchoprovocation with Ag. There was a striking reduction in mIL-5Ralpha on airway eosinophils compared with circulating cells. Furthermore, sIL-5Ralpha concentrations were elevated in BAL fluid, but steady state levels of sIL-5Ralpha mRNA were not increased in BAL compared with blood eosinophils. Finally, BAL eosinophils were refractory to IL-5 for ex vivo degranulation, suggesting that the reduction in mIL-5Ralpha on BAL eosinophils may regulate IL-5-mediated eosinophil functions. Together, the loss of mIL-5Ralpha, the presence of sIL-5Ralpha, and the blunted functional response (degranulation) of eosinophils to IL-5 suggest that when eosinophils are recruited to the airway, regulation of their functions becomes IL-5 independent. These observations provide a potential explanation for the inability of anti-IL-5 therapy to suppress airway hyperresponsiveness to inhaled Ag, despite a reduction in eosinophil recruitment.     

GM-CSF production by glioblastoma cells has a functional role in eosinophil survival, activation, and growth factor production for enhanced tumor cell proliferation

Medicinal interventions of limited efficacy are currently available for the treatment of glioblastoma multiforme (GBM), the most common and lethal primary brain tumor in adults. The eosinophil is a pivotal immune cell in the pathobiology of atopic disease that is also found to accumulate in certain tumor tissues. Inverse associations between atopy and GBM risk suggest that the eosinophil may play a functional role in certain tumor immune responses. To assess the potential interactions between eosinophils and GBM, we cultured human primary blood eosinophils with two separate human GBM-derived cell lines (A172, U87-MG) or conditioned media generated in the presence or absence of TNF-α. Results demonstrated differential eosinophil adhesion and increased survival in response to coculture with GBM cell lines. Eosinophil responses to GBM cell line-conditioned media included increased survival, activation, CD11b expression, and S100A9 release. Addition of GM-CSF neutralizing Abs to GBM cell cultures or conditioned media reduced eosinophil adhesion, survival, and activation, linking tumor cell-derived GM-CSF to the functions of eosinophils in the tumor microenvironment. Dexamethasone, which has been reported to inhibit eosinophil recruitment and shrink GBM lesions on contrast-enhanced scans, reduced the production of tumor cell-derived GM-CSF. Furthermore, culture of GBM cells in eosinophil-conditioned media increased tumor cell viability, and generation of eosinophil-conditioned media in the presence of GM-CSF enhanced the effect. These data support the idea of a paracrine loop between GM-CSF-producing tumors and eosinophil-derived growth factors in tumor promotion/progression.